Protective effect of polysaccharide nucleic acid fraction of bacillus Calmette-Guerin against atopic dermatitis in Nc/Nga mice / 中华皮肤科杂志

Objective To evaluate the protective effect of polysaccharide nucleic acid fraction of bacillus Calmette-Guerin (BCG-PSN) against atopic dermatitis (AD) in Nc/Nga mice,and to explore its possible mechanism.Methods Sixteen Nc/Nga mice were classified into normal control group (n =4),low-concentration BCG-PSN group (n=5) and high-concentration BCG-PSN group (n =7) to be subcutaneously injected with sodium chloride physiological solution,BCG-PSN of 0.1 mg/kg and 0.5 mg/kg respectively,at 1,8,15 and 22 days of age.Dinitrochlorobenzene (DNCB) was repeatedly and topically applied to these Nc/Nga mice to induce AD-like lesions at 49 days of age.The preventive effect of BCG-PSN against AD was evaluated by dermatitis scores,scratching frequency,histopathological manifestations and immunological parameters (including IgE,i nterleukin (IL)-4 and-12,and interferon (IFN)-γ).Results Repeated injection of BCG-PSN within 4 weeks after birth significantly decreased the severity of DNCB-induced AD-like lesions,dermatitis scores and scratching behavior in Nc/Nga mice.There was no statistical difference in scratching frequency between the high-and low-concentration BCG-PSN groups.BCG-PSN treatment reduced the plasma level of IgE in Nc/Nga mice in a dose-dependent manner.BCG-PSN at 0.5 mg/kg increased the number of cells secreting IFN-γ in skin lesions of mice.Both doses of BCG-PSN down-regulated IL-4 level,but up-regulated IL-12 level in the culture supernatant of spleen mononuclear cells from mice.Conclusion Early injection of BCG-PSN could protect Nc/Nga mice against dermatitis by promoting the proliferation of IFN-γ-secreting cells,increasing the synthesis of IL-12,and reducing the levels of IL-4 and IgE.

Clinical observations on the efficacy of total glucosides of paeony combined with ebastine in the treatment of chronic idiopathic urticaria / 中华皮肤科杂志

Objective To evaluate the efficacy and safety of total glucosides of paeony combined with ebastine for the treatment of chronic idiopathic urticaria.Methods A randomized,open-labeled,positive drugcontrolled,parallel-group study was carried out.Sixty patients with chronic idiopathic urticaria were randomly divided into two groups using a random digit table:combined group treated with total glucosides of paeony 600 mg thrice daily combined with ebastine 10 mg per day,control group treated with ebastine 10 mg per day only.The treatment lasted 12 weeks followed by a four-week follow-up.Adverse reactions were recorded and treatment efficacy was evaluated by using urticaria activity score over seven days (UAS7).Results The UAS7 was 9.28 ±4.59,5.83 ± 4.44 and 7.52 ± 5.57 in the combined group on week 8,12 after the initiation of treatment and week 4 after the withdrawal of treatment,respectively,significantly lower than that in the control group at the three time points (13.29 ± 4.72,P ＜ 0.05; 9.86 ± 5.46,P ＜ 0.01; 16.21 ± 5.34,P ＜ 0.01).Significant differences were observed in the response rate between the combined group and control group at the end of the 12-week treatment (75.9％ vs.42.9％,x2 =4.56,P ＜ 0.05).There was a decreased recurrence rate in the combined group combined with the control group at the end of the follow-up (13.6％ vs.50.0％,x2 =3.90,P ＜0.05).No obvious adverse reactions were noted in either of the two groups.Conclusion Total glucosides of paeony could markedly enhance the efficacy of ebastine for the treatment of chronic idiopathic urticaria with a reduction in recurrence rate.

Inhibitive effect of triptolide on invasiveness of human fibrosarcoma cells by downregulating matrix metalloproteinase-9 expression

OBJECTIVE@#To explore the molecular mechanisms of antitumor properties of triptolide, a bioactive component isolated from the Chinese herb Tripterygium wolfordii Hook F.@*METHODS@#Human fibrosarcoma HT-1080 cells were treated with different doses of triptolide for 72 h. Then the expression and activity of matrix metalloproteinase (MMP)-2 and -9 were measured and the invasiveness of triptolide-treated HT-1080 cells was compared with that of anti-MMP-9-treated HT-1080 cells.@*RESULTS@#18 nmol/L triptolide inhibited the gene expression and activity of MMP-9, but not those of MMP-2, in HT-1080 cells. In addition, both 18 nmol/L triptolide and 3 μg/mL anti-MMP-9 significantly reduced the invasive potential of HT-1080 cells, by about 50% and 35%, respectively, compared with the control. Whereas there was no significant difference between the effect of 18 nmol/L triptolide and that of anti-MMP-9 on invasive potential of HT-1080 cells.@*CONCLUSIONS@#These data suggest that triptolide inhibits tumor cell invasion partly by reducing MMP-9 gene expression and activity.

Effects of hypoxia inducible factor-1 α siRNA on inducible nitric oxide synthase expression in HaCaT cells / 中南大学学报(医学版)

OBJECTIVE@#To observe the effect of hypoxia inducible factor -1α (HIF-1α) small interfering RNA (siRNA) on the expression of HIF-1α and inducible nitric oxide synthase (iNOS) in HaCaT cells under hypoxia.@*METHODS@#HaCaT cells were divided into 4 groups: the normal control group (without any treatment), the hypoxia group (under hypoxia for 24 h), the liposome control group (HaCaT cells transfected with liposome before hypoxia treatment), the RNA interference group (HaCaT cells transfected with siRNA sequences then under hypoxia for 24 h). Real-time PCR and Western blot were utilized to determine HIF-1α and iNOS mRNA and protein expression in HaCaT cells.@*RESULTS@#There was no significant difference of the mRNA expression of HIF-1α between the hypoxia group and the normoxia group (P>0.05), but the protein expressions of HIF-1α was increased in the hypoxic group than that in the normoxia group (P<0.05). Both the mRNA and protein expression of iNOS were increased in hypoxic conditions than that in the normoxia (P<0.05). Decreases were more significant in the mRNA and protein expression of HIF-1α and iNOS in the RNA interference group than that in the liposome control group in HaCaT cells (P<0.05).@*CONCLUSION@#Hypoxia increased HIF-1α and iNOS expression in HaCaT cells and inhibition of HIF-1α expression decreased iNOS expression.

Effects of hypoxia inducible factor-1 alpha-targeting small interfering RNA on vascular endothelial growth factor gene expression in HaCaT cells / 中华皮肤科杂志

ObjectiveTo observe the effects of hypoxia inducible factor-1 alpha (HIF-1α)-targeting small interfering RNA(siRNA) on the expression of HIF-1α and vascular endothelial growth factor (VEGF) in HaCaT ceils under hypoxic conditions. MethodsHaCaT cells were cultured and divided into four groups, normal control group (without any treatment), hypoxia group (cultured under hypoxic conditions for 24 hours),liposome control group (transfected with liposome followed by hypoxic culture for 24 hours), RNA interference group (transfected with HIF-1α-targeting siRNA/liposome complexes followed by hypoxic culture for 24 hours). Fluorescence real-time quantitative PCR was utilized to determine HIF-1oα and VEGF mRNA expression in HaCaT cells, and Western blot to detect HIF-1α and VEGF protein expression. ResultsNo significant difference was observed in the mRNA expression of HIF-1α between the hypoxia group and normal control group(0.907 ± 0.032 vs. 0.878 ± 0.034, F =1.108, P ＞ 0.05), while the expression levels of VEGF mRNA,HIF-1α and VEGF proteins were significantly higher in the hypoxia group than in the normal control group (0.935 ± 0.032 vs. 0.652 ± 0.053, 0.813 ± 0.047 vs. 0.236 ± 0.014, 0.791 ± 0.030 vs. 0.316 ± 0.013, all P ＜0.05). A significant decline was noted in the mRNA and protein expression levels of VEGF (0.230 ± 0.044 vs.0.978 ± 0.030, 0.213 ± 0.026 vs. 0.817 ± 0.049, both P ＜ 0.05) and HIF-1α(0.497 ± 0.033 vs. 0.806 ±0.040, 0.249 ± 0.028 vs. 0.833 ± 0.052, both P ＜ 0.05) in the RNA interference group than in the liposome control group. ConclusionsHypoxia may enhance the expression of HIF-1α and VEGF in HaCaT cells, and to inhibit the HIF-1α expression may suppress the expression of VEGF in HaCaT cells under hypoxia.

Expression of iNOS and HIF-1α with angiogenesis in affected skin biopsies from patients with psoriasis / 中南大学学报(医学版)

OBJECTIVE@#To investigate the expression of inducible nitric oxide synthase (iNOS) and hypoxia-inducible factor 1(HIF-1) α in psoriatic lesions,and to explore their relationship with angiogenesis in psoriasis.@*METHODS@#HIF-1α and iNOS protein were detected by immunohistochemistry and Western blot in 32 cases of psoriasis and 20 healthy controls,and CD34 marking vascular endothelial cells were used to measure the microvascular density (MVD).@*RESULTS@#The expressions of HIF-1α and iNOS protein were very weak in the control skin, but very strong in psoriatic lesions, which showed significant difference in the expressions of iNOS and HIF-1α between the psoriasis and the control group(P<0.05). Expression of HIF-1α (r=0.743, P<0.01) and iNOS (r=0.639,P<0.01) had positive correlation with MVD in psoriasis respectively. There was a positive correlation between iNOS and HIF-1α expression in psoriasis (r=0.717, P<0.01).@*CONCLUSION@#Both iNOS and HIF-1α have high expression in psoriasis and might play an important role in the genesis and development of psoriasis.

Effects of BCG-PSN on 2,4-dinitrochlorobenzene-induced atopic dermatitis-like skin lesions in Nc/Nga mice / 中华皮肤科杂志

Objective To determine the effect of bacille Calmette-Guerin-polysaccharide nucleic acid (BCG/PSN)on 2,4-dinitrochlorobenzene(DNCB)-induced atopic dermatitis-like skin lesions in Nc/Nga mice.Methods Fifteen mice were randomly and equally classified into 3 groups,i.e.,control group receiving topical acetone on foot pad and abdomen and intraperitoneal injection of physiological saline,model group receiving topical 5% DNCB solution and intraperitoneal injection of physiological saline,treatment group receiving 5% DNCB solution and intraperitoneal iniection of BCG/PSN,and all drugs were used every other day for 7 weeks.Further more,0.1% DNCB was topically applied on the ear and neck of Nc/Nga mice once a week from week 2 to week 7.The effects of BCG/PSN were evaluated by ear thickness,skin histopathology and immunological parameters.Results Repeated application of DNCB caused the development of eczematous dermatitis in mice.Mice in model group chnieally manifested skin dryness,erythema,edema and erosion with histopathological changes including dermal and epidermal thickening,hyperkeratosis,and inflammatory infiltration.The serum levels of IL-4 and IrE in model group were significantly higher than those in control group[(174.72±12.64)μg/L vs (17.32±3.56)μg/L,(91.49±6.32)ng/L vs (83.95±6.63)ng/L,both P<0.05].Increased serum IL-12 and IFN-γ and decreased serum IgE were observed in treatment group compared with the model group[(122.10±4.64)ng/L vs (20.14±6.15)ng/L(73.89±2.39)ng/L vs (51.53±3.45)ng/L, (84.27±9.35)μg/L vs (174.72±12.64)μg/L, all P<0.05].Conclusion BCG/PSN might be beneficial for the treatment of atopie dermatitis-like skin lesions in Nc/Nga mice by enhancing the secretion of IL-12 and IFN-γ and suppressing the synthesis of IgE.

Triptolide evaluates DNA methylation level of matrix metalloproteinase 9 gene in human fibrosarcoma HT-1080 cells / 中国中药杂志

<p><b>OBJECTIVE</b>The effect of triptolide on the DNA methylation level of MMP-9 gene and the mRNA expression of tissue inhibitors of met-alloproteinases (TIMPs) were examined in human fibrosarcoma HT-1080 cells to explore the molecular mechanisms involved in the anticancer activity of triptolide.</p><p><b>METHOD</b>HT-1080 cells were cultured in MEM containing 10% newborn calf serum and 1% penicillin-streptomycin. Triptolide was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 1 goL-1 and stored at -20 degrees C. Triptolide was freshly diluted with culture medium perior to use and directly added to cell cultures at the indicated concentration, and incubated for 72 hours at 37 degrees C in a humidified atmosphere with 5% CO2, with changes of reagents every 24 hours. Methylation specific PCR (MSP)was applied to assess the methylation status of MMP-9 gene promoter, and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was employed to measure the mRNA expression of tissue inhibitors of metalloproteinases (TIMPs) in human fibrosarcoma HT-1080 cells after 72 hours of treatment with 6 nmol x L(-1), 12 nmol x L(-1) or 18 nmol x L(-1) triptolide, respectively.</p><p><b>RESULTS</b>The methylation index of MMP-9 gene promoter was statistically elevated in HT-1080 cells after 72 hours of treatment with 18 nmol L(-1) triptolide, compared with those in controls (0.61 +/- 0.10 vs 0.39 +/- 0.10, P < 0.05), while no significant difference was noted between 6 nmol x L(-1) or 12 nmol x L(-1) triptolide treated HT-1080 cells and controls (0.40 +/- 0.15 vs 0.39 +/- 0.10, 0.46 +/- 0.20 vs 0.39 +/- 0.10, respectively, both P > 0.05). The mRNA expression of TIMP-1, -2, -3 or -4 was not significantly changed in HT-1080 cells after 72 hours of treatment with the indicated concentrations of triptolide, respectively compared with those in controls (all P > 0.05).</p><p><b>CONCLUSION</b>The results demonstrated that triptolide upregulates the methylation level of MMP-9 gene in HT-1080 cells in vitro.</p>

Effect of phorbol-12-myristate-13-acetate on cyclooxygenase-2 expression in human HaCaT keratinocytes / 中华皮肤科杂志

Objective To investigate the effect of phorbol-12-myristate-13-acetate(PMA)on cyclooxygenase-2(COX-2) mRNA and protein expression in cultured human HaCaT keratinocytes,and the mechanism for cytotoxity of PMA against keratinocytes.MethodsRT-PCR and Westem blot were utilized to detect the expression of COX-2 mRNA and protein in cultured HaCaT ells at 24 hours after the treatment with various concentrations of PMA (0.1,1.0,10 mg/L).ResultsWithout any treatment,there was no or a weak expression df COX-2 mRNA and protein in HaCaT cells;incubation witll PMA resulted in the induction of the expression of COX-2 in HaCaT cells.The expression levels of COX-2 mRNA and protein in 10 mg/L PMA-pretreated HaCaT cells were significantly higher than those in 1.0 mg/L PMA-pretreated HaCaT cells,which was in turn higher than that in 0.1 mg/L PMA-pretreated cells and untreated cells;the difrerence was statistically significant (all P＜0.01).Conclusion These results suggest that PMA may be involved in keratinocyte tumorigenesis by upregulating he expression of COX-2 as well as synthesis and release of prostaglandin in keratinocytes.

Test of Urogenital Tract Infection With Chlamydiae Trachomatis and Mycoplasm and Drug Susceptibility Analysis in 5095 Cases / 中国医师杂志

Objective To investigate the infection of chlamydiae trachomahs(CT) and myoplasmas in urogenital tract and antibiotic susceptibility of cultured genital myoplasmas.Methods 5095 patients with urogenital tract infection were detected with mycoplasma identification susceptibility testing reagent kit,and the drug susceptibility to eight antibiotics of ureaplasma urealyticum (Uu) and mycoplasm hominid (Mh) were tested by broth microdilution method.And chlamydiae trachomatis was examined by golden standard method.Results In 5095 cases, 417cases(8 2%) were infected with chlamydiae trachomatis, and the infective rate in woman(11 2%) was statistically higher than that in man(6 2%). 1728 cases(33 9%) were infected with mycoplasma, and the infective rate in woman(43 0%) was statistically higher than that in man(28 0%).The cases infected with simple UU(1251,24 6%) were more than that in the cases infected with simple Mh(71,1 4%) and the mixed infected cases(406,8 0%). Drug sensitivity to erythromycin, roxithromycin, josamycin, azithromycin, doxycycline, minocin,ofloxacin, ciprofloxacin in Uu infection were 60 3%,67 5%,73 2%,85 3%,55 7%,40 1%,25 6%,2 7%,respectively;while the mixed infection of Mh and Uu had resistance to the eight antibiotics on the different degree.Conclusions The infective rate of chlamydiae trachomatis and myoplasma in urogential tract and the resistance rates to 8 antibiotics in Hunan province were in a higher level, compared with other area inland. It is necessary to develop antibiotic susceptibility test,in addition to the myoplasma culture for guiding the clinical therapy.

Expression of Inducible Nitric Oxide Synthase and NF-κ Bp65 in Patients with Psoriasis Vulgaris and Their Clinical Significance / 中华皮肤科杂志

Objective To investigate the internal relationship between the expression of inducible nitric oxide synthase (iNOS) and the activation of NF-κ Bp65 in the skin of patients with psoriasis vulgaris(PV) and their roles in the pathogenesis of PV.Methods The expression and distribution of iNOS and NF-κ Bp65 were studied by in situ hybridization and immunohistochemistry in 20 patients with PV and 12 healthy controls.The severity of PV was assessed by Psoriasis and Severity Index(PASI) score.Results ① The expression of both iNOS mRNA and protein was significantly higher in lesional epidermis than those in non-lesional epidermis and epidermis of the controls (P<0.01).Weak exprssion of iNOS mRNA was found in basal cell layer in a small number of healthy controls, however, strong expression was observed in entire basal cell layer and focal parts of prickle cell layer in all PV epidermis.② There was significantly positive correlation between iNOS and NF-κ Bp65 protein expression in lesional epidermis (P< 0.01).③ There was significantly positive correlation between the level of expression of iNOS mRNA in lesional epidermis and PASI score of patients (P< 0.01).Conclusions The expression of iNOS and NF-κ Bp65 is markedly increased in lesional epidermis of patient with PV.The activation of NF-κ B probably takes part in the pathogenesis of PV.High-level expression of iNOS may be due to the activation of NF-κ B.

The effect of RNA interference on the expression of CTGF in skin fibroblasts of systemic sclerosis / 中华皮肤科杂志

Objective To study the effect of RNA interference on the expression of CTGF in skin fibroblasts of systemic sclerosis(SS). Methods Four CTGF specific siRNAs and a negative control siRNA were designed and then synthesized by in vitro transcription. siRNAs labeled with carboxyfluorescein-6-succimidyl ester (FAM) were transiently transfected into SS skin fibroblasts. Forty-eight hours after the fibroblasts were treated with siRNAs, the mRNA and protein expression of CTGF was detected by semiquantitative RT-PCR and Western blot analysis, respectively. Results The mRNA and protein expression of CTGF in fibroblasts was significantly down-regulated by 4 and 3 CTGF specific siRNAs (both P

Expression of Inducible Nitric Oxide Synthase and NF－? Bp65 in Patients with P soriasis Vulgaris and Their Clinical Significance / 中华皮肤科杂志

Objective To investigate the internal relationship between the exp ression of inducible nitric oxide synthase (iNOS) and the activation of NF－? B p65 in the skin of patients with psoriasis vulgaris(PV) and their roles in the p athogenesis of PV. Methods The expression and distribution of iNOS and NF－? Bp 65 were studied by in situ hybridization and immunohistochemistry in 20 patients with PV and 12 healthy controls. The severity of PV was assessed by Psoriasis a nd Severity Index(PASI) score. Results① The expression of both iNOS mRNA and pr otein was significantly higher in lesional epidermis than those in non lesiona l epidermis and epidermis of the controls (P